Mycobacterium tuberculosis Nucleic Acid Detection Reagent
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Solution Overview
Problem
Current laboratory diagnosis methods for tuberculosis are inefficient, time-consuming, and struggle to distinguish between latent and active infections, and lack effective monitoring indicators for treatment, particularly with the challenges of multidrug-resistant TB and co-infection with HIV.
Innovation Solution
A nucleic acid detection reagent and method for Mycobacterium tuberculosis using Tris-HCL, MgCl2, UNG enzyme, Bst DNA polymerase, reverse transcriptase, trehalose, sucrose, mannitol, Tween 20, guanidine hydrochloride, and specific primers for one-step detection of DNA or RNA in throat swabs, nasal swabs, and saliva, enabling rapid and convenient detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional acid-fast staining and culture methods are used for tuberculosis diagnosis, then the diagnostic process can be performed with simple equipment, but the detection positive rate is low, specificity is poor, and the time required is long
Solution Approach 1:
The patent replaces traditional mechanical/culture-based diagnosis methods with a nucleic acid detection system that uses enzymatic reactions (reverse transcriptase and DNA polymerase) to amplify and detect Mycobacterium tuberculosis DNA or RNA, achieving rapid and accurate detection without requiring lengthy culture periods
Solution Approach 2:
The patent changes the detection parameter from physical/cultural characteristics to molecular nucleic acid sequences, using specific primer pairs (TB-F3/TB-R3, TB-FIP/TB-BIP, TB-LF/TB-LB) to target and detect specific genetic markers of M. tuberculosis, thereby improving both speed and accuracy
2Measurement precision
If traditional laboratory diagnosis methods are used, then the equipment requirements are simple, but the ability to distinguish latent infection from active tuberculosis is insufficient
Solution Approach 1:
The patent uses different primer pairs to detect different nucleic acid markers: TB-F3/TB-R3 and TB-FIP/TB-BIP for active tuberculosis detection, while TB-LF/TB-LB targets latent infection markers, enabling differentiation between latent and active states through molecular parameter analysis
Solution Approach 2:
The patent segments the detection system into multiple independent primer pairs, each targeting specific regions of the M. tuberculosis genome associated with different infection states, allowing simultaneous detection of multiple conditions using a single integrated reagent system
3Ease of operation
If conventional detection methods are used, then the procedure is straightforward, but there are no effective indicators for monitoring tuberculosis treatment
Solution Approach 1:
The patent provides quantitative nucleic acid detection results that can be used to monitor treatment response over time, allowing clinicians to assess whether patient nucleic acid loads are decreasing as expected during therapy, thereby establishing a feedback mechanism for treatment monitoring
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The solution provides a rapid, efficient, and convenient detection method for Mycobacterium tuberculosis, eliminating the need for nucleic acid extraction and allowing for direct visualization of results, suitable for various application scenarios including clinics and home testing, with high sensitivity and specificity.
Implementation Method 1
comprising: Tris-HCL, MgCl2, UNG enzyme, Bst DNA polymerase, reverse transcriptase
Data Source
AI summary
The present application provides a nucleic acid detection reagent and detection method for Mycobacterium tuberculosis, comprising: Tris-HCL, MgCl2, UNG enzyme, Bst DNA polymerase, reverse transcriptase, trehalose, sucrose, mannitol, Tween 20, guanidine hydrochloride and the primers of six kinds of Mycobacterium tuberculosis, the primers of six kinds of Mycobacterium tuberculosis are TB-F3, TB-R3, TB-FIP, TB-BIP, TB-LF and TB-LB, wherein TB is tuberculosis branch bacilli. In the embodiment of the present application, the nucleic acid detection reagent can be used to perform a one-step detection method for the nucleic acid (DNA or RNA) of pathogens present in throat swabs, nasal swabs, saliva, etc., thereby realizing a convenient, efficient and rapid detection, to achieve the sample in, the result out.